Polypeptides having phospholipase C activity and polynucleotides encoding same

Abstract

The present invention relates to a method of reducing the phospholipid content in an oil or fat composition and polypeptides having PI-specific phospholipase C activity as well as polypeptides having PC, PE-specific phospholipase C activity and combinations thereof capable of catalyzing this reduction. The invention also relates to polynucleotides encoding the polypeptides, nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

The invention claimed is: 
     
         1 . A composition comprising
 (a) a phosphatidylinositol phospholipase C having at least 90% sequence identity to amino acid residues 26-322 of SEQ ID NO: 2 or amino acid residues 1-298 of SEQ ID NO: 3; and   (b) a phosphatidylcholine and phosphatidylethanolamine-specific phospholipase C polypeptide (“a PC and PE-specific phospholipase C”) having at least 90% sequence identity to amino acid residues 25-280 of SEQ ID NO: 38 or amino acid residues 36-280 of SEQ ID NO: 38.   
     
     
         2 . The composition of  claim 1 , wherein
 (a) the phosphatidylinositol phospholipase C has at least 95% sequence identity to amino acid residues 26-322 of SEQ ID NO: 2 or amino acid residues 1-298 of SEQ ID NO: 3.   
     
     
         3 . The composition of  claim 1 , wherein
 (b) the PC and PE-specific phospholipase C has at least 95% sequence identity to amino acid residues 25-280 of SEQ ID NO: 38 or amino acid residues 36-280 of SEQ ID NO: 38.   
     
     
         4 . The composition of  claim 1 , wherein
 (a) the phosphatidylinositol phospholipase C has at least 95% sequence identity to amino acid residues 26-322 of SEQ ID NO: 2 or amino acid residues 1-298 of SEQ ID NO: 3; and   (b) the PC and PE-specific phospholipase C has at least 95% sequence identity to amino acid residues 25-280 of SEQ ID NO: 38 or amino acid residues 36-280 of SEQ ID NO: 38.   
     
     
         5 . The composition of  claim 1 , wherein
 (a) the phosphatidylinositol phospholipase C has at least 98% sequence identity to amino acid residues 26-322 of SEQ ID NO: 2 or amino acid residues 1-298 of SEQ ID NO: 3.   
     
     
         6 . The composition of  claim 1 , wherein
 (b) the PC and PE-specific phospholipase C has at least 98% sequence identity to amino acid residues 25-280 of SEQ ID NO: 38 or amino acid residues 36-280 of SEQ ID NO: 38.   
     
     
         7 . The composition of  claim 1 , wherein
 (a) the phosphatidylinositol phospholipase C has at least 98% sequence identity to amino acid residues 26-322 of SEQ ID NO: 2 or amino acid residues 1-298 of SEQ ID NO: 3; and   (b) the PC and PE-specific phospholipase C has at least 98% sequence identity to amino acid residues 25-280 of SEQ ID NO: 38 or amino acid residues 36-280 of SEQ ID NO: 38.   
     
     
         8 . The composition of  claim 1 , wherein
 (a) the phosphatidylinositol phospholipase C comprises amino acid residues 26-322 of SEQ ID NO: 2 or amino acid residues 1-298 of SEQ ID NO: 3.   
     
     
         9 . The composition of  claim 1 , wherein
 (b) the PC and PE-specific phospholipase C polypeptide comprises amino acid residues 25-280 of SEQ ID NO: 38 or amino acid residues 36-280 of SEQ ID NO: 38.   
     
     
         10 . The composition of  claim 1 , wherein
 (a) the phosphatidylinositol phospholipase C comprises amino acid residues 26-322 of SEQ ID NO: 2 or amino acid residues 1-298 of SEQ ID NO: 3; and   (b) the PC and PE-specific phospholipase C polypeptide comprises amino acid residues 25-280 of SEQ ID NO: 38 or amino acid residues 36-280 of SEQ ID NO: 38.   
     
     
         11 . The composition of  claim 1 , wherein
 (a) the phosphatidylinositol phospholipase C is a fragment of amino acid residues 26-322 of SEQ ID NO: 2 or a fragment of amino acid residues 1-298 of SEQ ID NO: 3.   
     
     
         12 . The composition of  claim 1 , wherein
 (b) the phosphatidylcholine (“PC”) and phosphatidylethanolamine (“PE”)-specific phospholipase C polypeptide is a fragment of amino acid residues 39-283 of SEQ ID NO: 22.   
     
     
         13 . The composition of  claim 1 , wherein
 (a) the phosphatidylinositol phospholipase C is a fragment of amino acid residues 26-322 of SEQ ID NO: 2 or a fragment of amino acid residues 1-298 of SEQ ID NO: 3; and   (b) the phosphatidylcholine (“PC”) and phosphatidylethanolamine (“PE”)-specific phospholipase C polypeptide is a fragment of amino acid residues 39-283 of SEQ ID NO: 22.